目的:探讨亚硫酸钠引起肝细胞内活性氧水平变化与p53蛋白表达之间的关系.方法；将人正常二倍体肝细胞株HL-7702培养于含10％小牛血清的DMEM培养基中,待生长至适宜密度时用不同浓度亚硫酸钠染毒4h.采用Fluo-3 AM钙离子荧光指示剂法测定肝细胞内Ca2+水平变化,二氯荧光黄双乙酸酯荧光标记法测定肝细胞内活性氧水平,蛋白杂交法和免疫细胞化学法检测肝细胞内p53蛋白表达情况,RT-PCR法检测肝细胞内p53基因mRNA表达水平的变化.结果:一定浓度的亚硫酸钠可引起肝细胞内Ca2+水平增加,并伴随着活性氧水平增加和肝细胞内p53蛋白表达水平明显增加,但p53基因mRNA表达水平无明显变化.结论:亚硫酸钠可通过增加肝细胞内Ca2+水平引起肝细胞内活性氧水平和p53蛋白水平增加,这种影响可能发生在蛋白质翻译或翻译后水平.%Objective:To study the possible relationship between reactive oxygen species (ROS) production and p53 protein expression of hepatocytes induced by sodium sulfite. Methods:The HL-7702 hepatocytes were cultured in DMEM medium containing 10% new born bovine serum. After 4h, intracellular calcium level was demonstrated using Fluo-3 acetome-thoxyester and the generation of ROS was probed with 2' ,7′ -dichlorofluorescein diacetate (DCFH-DA) fluorescence. The expression of p53 protein was detected by immunohistochemistry and Western blotting. The mRNA expression of p53 was demonstrated by RT-PCR. Results:Na2 SO3 increased Ca2+ level inside hepatocytes with an increase of ROS. The expression of p53 protein was also promoted by Na2SO3 treatment, and no significant changes in mRNA level of p53 were found. Conclusion:Na2SO3 can promote ROS production with an increase in p53 protein expression, which may be due to the increase of Ca2+ , and result in liver injury.
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